The long-term objective of this STTR project is to commercialize the protein multi-blot Western device, which contributes to improving the productivity and accuracy of conventional Western blot analysis. Although Western blotting has been used for more than three decades, this labor-and-cost demanding technique remains a key routine for a majority of life scientists. To improve Western blot analysis, new products for quick antibody reactions, better quantitation through imaging and software, and capillary electrophoresis for fully automation have been released. However, few products are directly targeted to improve a traditional blotting process which is still widely used. In this project, YC Bioelectric, LLC (YCB) proposes to develop a novel multi-blot Western device, which enables multiple copies of protein species of varying sizes from a single electrophoretic gel. In this device, a protein transfer unit utilizes multiple anodes to generate a non-uniform voltage profile in order to enable uniform protein transfer speed regardless of protein size. A preliminary study was conducted at Indiana University, and results show that the expression levels different proteins are detectable from each of 5-7 membranes made from a single gel. YCB was founded in June 2012 to manufacture a marketable multi-blot Western device product. YCB currently holds an option to take an exclusive license from the Indiana University Research and Technology Commercialization. YCB and Indiana University propose to extend the current proof-of-concept investigation to a full feasibility study. The hypothesis is that multi-blotting utilizing a multi-anode plate not only enhances accuracy of Western blot analysis but also significantly reduces cost and labor. The hypothesis will be tested with the following specific aims: Aim 1: Fabrication of the refined blotting hardware; Aim 2: Implementation of the enhanced control system; Aim 3: Development of a predictive software tool The revised proposal includes the development of a novel one button operated imaging based blotting control subsystem and an inclusion of a predictive software tool. Furthermore, solutions to four technical weaknesses in the last review regarding a voltage gradient, anode materials, quality control, and gel handling, are presented. According to its market analysis, YCB expects that its target customers in life science labs will appreciate the proposed multi-blot Western device because of its capability of saving costs, time, and samples with a user-friendly predictive software tool.